Multiple myeloma treatment

ABSTRACT

The invention relates generally to the treatment of multiple myeloma. One embodiment of the invention provides a method of treating multiple myeloma (MM) in an individual, the method comprising: administering to the individual an effective amount of trichostatin A (TSA).

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of co-pending U.S. ProvisionalPatent Application Nos. 61/869,039, filed 22 Aug. 2013 and 61/870,747,filed 27 Aug. 2013, each of which is incorporated herein.

BACKGROUND

Multiple myeloma (MM), sometimes referred to as plasma cell myeloma, isa multifocal plasma cell cancer of the osseous system, generallyaffecting elderly individuals. Most individuals are symptomatic whendiagnosed, with diagnosis typically made by one or more of serum proteinelectrophoresis, serum free kappa/lambda light chain assay, urineprotein electrophoresis (99% of patients with MM exhibit increasedlevels of one of the immunoglobulin (Ig) classes in the blood and/orlight chains in the urine), bone marrow examination, or X-ray analysis.Although MM generally responds to chemotherapy, recurrence is common,since such treatment does not target cancer stem cells.

Nara et al. have recently identified a number of candidate genes fortargeting MM tumor-initiating subpopulation (SP) cells, i.e., cancerstem cells. These include a number of genes coding for proteinsassociated with cell cycle and mitosis, all of which were found to beupregulated in MM cells. These include cyclin B1 (CCNB1), cell divisioncycle 2 (CDC2), baculoviral IAP repeat-containing 5 (BIRC5), abnormalspindle homolog, microcephaly-associated (ASPM), topoisomerase (DNA) IIalpha 170kDa (TOP2A), aurora kinase B (AURKB), kinesin family member 11(KIF11), and kinesin family member 2c (KIF2C).

Similarly, Shaughnessy et al. report a 70-gene high-risk profile formultiple myeloma. Two of the genes upregulated in this high-risk profileare CDC28 protein kinase regulatory subunit 1B (CKS1B) and WEE1 homolog(S. pombe) (WEE1).

SUMMARY

One embodiment of the invention provides a method of treating multiplemyeloma (MM) in an individual, the method comprising: administering tothe individual an effective amount of trichostatin A (TSA).

In another embodiment, the invention provides a method of treatingmultiple myeloma (MM) in an individual, the method comprising:determining, from a biological sample obtained from the individual'sbody, a level of expression of at least one gene selected from a groupconsisting of: CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM,CKS1B, and WEE1; and in the case that the level of expression of the atleast one gene is indicative of overexpression, administering to theindividual an effective amount of trichostatin A (TSA).

In yet another embodiment, the invention provides a method of treatingmultiple myeloma (MM) in an individual, the method comprising:diagnosing or having diagnosed the individual with MM; and administeringto the individual an effective amount of trichostatin A (TSA).

In still yet another embodiment, the invention provides a pharmaceuticalcomposition comprising: trichostatin A (TSA) as a sole or primaryinhibitor of CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM,CKS1B, or WEE1; and a pharmaceutically-acceptable excipient or carrier.

In still other embodiments of the invention, treatment with TSA iscombined with one or more other multiple myeloma treatments. Such othertreatments may include, for example, small molecule inhibition.

DETAILED DESCRIPTION

Trichostatin A (TSA or7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienamide),is an antifungal antibiotic. The structure of TSA is shown in Formula Ibelow.

Applicants have surprisingly found that TSA, although previously knownas a class I and II histone deacetylase (HDAC) inhibitor, is alsocapable of inhibiting expression of CCNB1, AURKB, CDC2, BIRC5, KIF11,KIF2C, TOP2A, ASPM, CKS1B, and WEE1. Accordingly, TSA may be used as aprimary or sole inhibitor of one or more such genes in the treatment ofMM.

A human retinal pigment epithelial cell line was treated withtrichostatin or vehicle for 24 hours and gene expression for 22,238probe sets covering 12,490 genes was generated using an Affymetrixinstrument. The effect of trichostatin A on expression of CCNB1, AURKB,CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1 is shown belowin Table 1, and indicates significant downregulation of the expressionof each gene.

TABLE 1 Instance ID Probe Rank Fold Expression Δ Gene 10005542219918_s_at 22283 −69.97232079 ASPM 10005533 219918_s_at 22282−54.61735261 ASPM 10005532 219918_s_at 22261 −23.24977266 ASPM 10005542209464_at 22190 −11.52858083 AURKB 10005533 209464_at 22185 −11.04347695AURKB 10005542 202095_s_at 22270 −24.2000252 BIRC5 10005533 202095_s_at22256 −23.02258123 BIRC5 10005533 202094_at 22251 −20.74385736 BIRC510005532 202095_s_at 22252 −19.95557418 BIRC5 10005542 202094_at 22227−14.71770993 BIRC5 10005532 202094_at 22219 −14.42912247 BIRC5 10005533214710_s_at 22267 −26.45555632 CCNB1 10005532 214710_s_at 22267−26.32053821 CCNB1 10005542 214710_s_at 22251 −20.15506664 CCNB110005532 203213_at 22270 −27.14720991 CDC2 10005533 203213_at 22260−23.81235655 CDC2 10005542 203213_at 22253 −20.26528442 CDC2 10005533210559_s_at 22199 −12.07146825 CDC2 10005532 210559_s_at 22192−11.92448867 CDC2 10005533 203214_x_at 22194 −11.8262682 CDC2 10005542204444_at 22213 −13.12379506 KIF11 10005532 204444_at 22187 −11.4579544KIF11 10005533 204444_at 22184 −10.96422696 KIF11 10005533 209408_at22250 −19.89427497 KIF2C 10005532 209408_at 22248 −19.35105571 KIF2C10005542 209408_at 22224 −14.47328923 KIF2C 10005532 201292_at 22274−31.9462153 TOP2A 10005533 201291_s_at 22270 −28.21627346 TOP2A 10005532201897_s_at 22279 −39.94584911 CKS1B 10005533 201897_s_at 22279−52.93016044 CKS1B 10005542 201897_s_at 22268 −23.90194858 CKS1B10005532 212533_at 22237 −17.0758281 WEE1 10005533 212533_at 22248−19.46663938 WEE1 10005542 212533_at 22265 −23.63054187 WEE1

These results support the use of TSA in the treatment of MM. Forexample, an individual may be treated for MM by administering to theindividual an effective amount of TSA, wherein the effective amount isan amount sufficient to inhibit expression of one or more of CCNB1,AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1 in theindividual. Such an amount may also be sufficient to inhibit HDACactivity in the individual. In some embodiments of the invention, theeffective amount is between about 0.01 mg/kg/day and about 100mg/kg/day, e.g., between about 0.1 mg/kg/day and about 10 mg/kg/day orbetween about 0.5 mg/kg/day and about 5 mg/kg/day.

In some embodiments, treating the individual may further comprisedetermining, from a biological sample obtained from the individual'sbody, a level of expression of one or more of CCNB1, AURKB, CDC2, BIRC5,KIF11, KIF2C, TOP2A, ASPM, CKS1B, or WEE1. Such determining may includeany known or later-developed method or technique, including, forexample, quantitative antigen-antibody interactions, the use of labelednucleotide probes, etc.

In other embodiments of the invention, treating the individual mayinclude diagnosing or having diagnosed the individual with MM prior toadministering TSA to the individual. Such diagnosing may include one ormore technique or method for making such a diagnosis, including, forexample, serum protein electrophoresis, serum free kappa/lambda lightchain assay, urine protein electrophoresis, bone marrow examination, orX-ray analysis.

TSA may be administered to the individual to be treated in the form of apharmaceutical composition. Pharmaceutical compositions to be usedaccording to various embodiments of the invention comprise atherapeutically effective amount of TSA or an active metabolite of TSA,or a pharmaceutically acceptable salt or other form (e.g., a solvate)thereof, together with one or more pharmaceutically acceptableexcipients or carriers. The phrase “pharmaceutical composition” refersto a composition suitable for administration in medical use. It shouldbe appreciated that the determinations of proper dosage forms, dosageamounts, and routes of administration for a particular patient arewithin the level of ordinary skill in the pharmaceutical and medicalarts.

Administration may be oral but other routes of administration may alsobe employed, e.g., parenteral, nasal, buccal, transdermal, sublingual,intramuscular, intravenous, rectal, vaginal, etc. Solid dosage forms fororal administration include capsules, tablets, pills, powders, andgranules. In such solid dosage forms, the compound is admixed with atleast one inert pharmaceutically-acceptable excipient such as (a)fillers or extenders, as for example, starches, lactose, sucrose,glucose, mannitol, and silicic acid, (b) binders, as for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone,sucrose, and acacia, (c) humectants, as for example, glycerol, (d)disintegrating agents, as for example, agar-agar, calcium carbonate,potato or tapioca starch, alginic acid, certain complex silicates, andsodium carbonate, (e) solution retarders, as for example paraffin, (f)absorption accelerators, as for example, quaternary ammonium compounds,(g) wetting agents, as for example, cetyl alcohol, and glycerolmonostearate, (h) adsorbents, as for example, kaolin and bentonite, and(i) lubricants, as for example, talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, or mixturesthereof. In the case of capsules, tablets, and pills, the dosage formsmay also comprise buffering agents. Solid dosage forms such as tablets,drages, capsules, pills, and granules also can be prepared with coatingsand shells, such as enteric coatings and others well known in the art.The solid dosage form also may contain opacifying agents, and can alsobe of such composition that they release the active compound orcompounds in a certain part of the intestinal tract in a delayed manner.Examples of embedding compositions which can be used are polymericsubstances and waxes. The active compounds can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-mentioned excipients. Such solid dosage forms may generallycontain from 1% to 95% (w/w) of the active compound. In certainembodiments, the active compound ranges from 5% to 70% (w/w).

Solid compositions for oral administration can be formulated in a unitdosage form, each dosage containing from about 0.1 mg to about 5000 mgof active ingredient. The term “unit dosage form” refers to physicallydiscrete units suitable as unitary dosages for human subjects and othermammals, each unit containing a predetermined quantity of activeingredient calculated to produce the desired effect over the course of atreatment period, in association with the required pharmaceuticalcarrier. TSA can be formulated, e.g., in a unit dosage form that is acapsule having 0.1-5000 mg of active in addition to excipients.

Liquid dosage forms for oral administration includepharmaceutically-acceptable emulsions, solutions, suspensions, syrups,and elixirs. In addition to the compound or composition, the liquiddosage forms may contain inert diluents commonly used in the art, suchas water or other solvents, solubilizing agents and emulsifiers, as forexample, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propyleneglycol,1,3-butyleneglycol, dimethylformamide, oils, in particular, cottonseedoil, groundnut oil, corn germ oil, olive oil, castor oil and sesame oil,glycerol, tetrahydrofurfuryl alcohol, polyethyleneglycols and fatty acidesters of sorbitan or mixtures of these substances. Besides such inertdiluents, the composition can also include adjuvants, such as wettingagents, emulsifying and suspending agents, sweetening, flavoring, andperfuming agents.

In some embodiments of the invention, TSA is provided in a liquid formand administered to an individual intravenously. According to someembodiments of the invention, TSA is provided in a sustained orcontrolled release formulation.

While this invention has been described in conjunction with the specificembodiments outlined above, it is evident that many alternatives,modifications and variations will be apparent to those skilled in theart or are otherwise intended to be embraced. Accordingly, theembodiments of the invention as set forth above are intended to beillustrative, not limiting. Various changes may be made withoutdeparting from the spirit and scope of the invention as defined in thefollowing claims. All patents, patent application, scientific articlesand other published documents cited herein are hereby incorporated intheir entirety for the substance of their disclosures.

1. A method of treating multiple myeloma (MM) in an individual, themethod comprising: administering to the individual an effective amountof trichostatin A (TSA).
 2. The method of claim 1, wherein the effectiveamount is an amount sufficient to decrease expression of at least onegene in the individual, the at least one gene being selected from agroup consisting of: CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A,ASPM, CKS1B, and WEE1.
 3. The method of claim 2, wherein the effectiveamount is an amount sufficient to decrease expression of either or bothof CCNB1 and AURKB in the individual.
 4. The method of claim 2, whereinthe effective amount is an amount sufficient to decrease expression ofeither or both of CKS1B and WEE1.
 5. The method of claim 2, wherein theeffective amount is an amount sufficient to decrease expression of eachof CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, andWEE1.
 6. The method of claim 1, wherein the effective amount is betweenabout 0.01 mg/kg/day and about 100 mg/kg/day.
 7. The method of claim 1,wherein the effective amount is between about 0.1 mg/kg/day and about 10mg/kg/day.
 8. The method of claim 7, wherein the effective amount isbetween about 0.5 mg/kg/day and about 5 mg/kg/day.
 9. The method ofclaim 1, wherein administering includes orally administering.
 10. Themethod of claim 1, wherein administering includes intravenouslyadministering.
 11. A method of treating multiple myeloma (MM) in anindividual, the method comprising: determining, from a biological sampleobtained from the individual's body, a level of expression of one ormore gene in the individual, the one or more gene selected from a groupconsisting of: CCNB1, AURKB, CDC2, BIRC5, KIF11, KIF2C, TOP2A, ASPM,CKS1B, and WEE1; and in the case that the level of expression of the oneor more gene is indicative of overexpression, administering to theindividual an effective amount of trichostatin A (TSA).
 12. The methodof claim 11, wherein the effective amount is an amount sufficient todecrease expression of at least one gene in the individual, the at leastone gene being selected from a group consisting of: CCNB1, AURKB, CDC2,BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1.
 13. The method ofclaim 12, wherein the effective amount is an amount sufficient todecrease expression of either or both of CCNB1 and AURKB in theindividual.
 14. The method of claim 12, wherein the effective amount isan amount sufficient to decrease expression of either or both of CKS1Band WEE1.
 15. The method of claim 12, wherein the effective amount is anamount sufficient to decrease expression of each of CCNB1, AURKB, CDC2,BIRC5, KIF11, KIF2C, TOP2A, ASPM, CKS1B, and WEE1.
 16. The method ofclaim 11, wherein the effective amount is between about 0.01 mg/kg/dayand about 100 mg/kg/day.
 17. The method of claim 16, wherein theeffective amount is between about 0.1 mg/kg/day and about 10 mg/kg/day.18. The method of claim 17, wherein the effective amount is betweenabout 0.5 mg/kg/day and about 5 mg/kg/day.
 19. The method of claim 11,wherein administering includes orally administering.
 20. The method ofclaim 11, wherein administering includes intravenously administering.21-42. (canceled)